NEST 50 mL Centrifuge Tube (500 count)

Biotin-PEG4-C1-alkyne is azide reactive biotinylation reagent, PEG4 arm increase aqueous solubility of the reagent. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Biotin-PEG4-C1-alkyne is azide reactive biotinylation reagent, PEG4 arm increase aqueous solubility of the reagent. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Overview

• Isolate all sizes of circulating and exosomal RNA, including microRNA
• Versatile urine input ranges
• No phenol extractions
• No carrier RNA
• Bind and elute all RNA irrespective of size or GC content, without bias
• Concentrate circulating RNA and exosomal RNA into flexible elution volumes
• Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating RNA, including exosomal RNA as well as viral RNA from fresh, preserved or frozen urine samples. All components for the purification are provided in one convenient and fast kit for the easy processing of small input volumes of bodily fluids. The purified urine RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.

Background

Recent evidence indicates that cell-free circulating RNA (cf-RNA) including exosomal RNA in urine contains valuable information for the discovery of biomarkers that can help for the early detection of certain cancer types and for monitoring the disease status as well as for the detection of any infectious pathogens. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. For this reason exosomal RNAs may serve as biomarkers for various diseases including cancer. The advantage for using urine as a source for cancer biomarkers is that it can be acquired in large quantities without using invasive procedures. In addition, repeated sampling from the same individual is applicable, which facilitates longitudinal studies. There are many advantages favouring the use of urinary nucleic acid for cancer biomarker discovery over blood, tissue samples or other bodily fluids, including: (1) urine is non-infectious for HIV and less infectious for many other pathogens; (2) the profile of urinary nucleic acid is similar to that in plasma or serum but with a lower concentration; (3) Nucleic acid purification from urine is technically much easier because of its low protein concentration (1000-fold lower than blood).

Urine Cell-Free Circulating RNA Purification Mini Kit

For sample volumes ranging from 250 µL to 2 mL.

Urine Cell-Free Circulating RNA Purification Midi Kit

For sample volumes ranging from 2 mL to 10 mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.

Urine Cell-Free Circulating RNA Purification Maxi Kit

For sample volumes ranging from 10 mL to 30 mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL

Details

Supporting Data

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Kit Specifications
Minimum Urine Input	10 mL
Maximum Urine Input	30 mL
Size of RNA Purified	All sizes including miRNA and small RNA (< 200 nt)
Elution Volume	50-100 μL
Time to Complete 10 Purifications	40-45 minutes
Average Yields	Variable depending on specimen

*Please check page 6 of the product insert for average yields and common RNA quantification methods.

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. These kits are stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

Component	Cat. 56900 (50 preps)	Cat. 57000 (20 preps)	Cat. 57100 (10 preps)
Binding Solution K	25 mL	75 mL	1 x 75 mL 1 x 25 mL
Lysis Buffer A	30 mL	20 mL	20 mL
Wash Solution A	18 mL	18 mL	18 mL
Elution Solution A	6 mL	6 mL	6 mL
Mini Spin Columns	50	20	10
Midi Spin Columns	–	20	–
Maxi Spin Columns	–	–	10
Collection Tubes	50	20	10
Elution Tubes (1.7 mL)	50	20	10
Product Insert	1	1	1

Introduction

Usages:
For differentiating enteric bacteria based on urease activity by adding 40% sterile urea solution.

Principle:
Peptone provides the carbon and nitrogen; maintain a balanced osmotic sodium chloride; potassium dihydrogen phosphate is buffers; decomposing bacteria urease urea medium, produce large amounts of ammonia, agar as medium coagulant.

Formulation (per liter):
Peptone 1g
Sodium chloride 5g
Glucose 1g
Ppotassium dihydrogen phosphate 2g
Phenol red 0.012g
Agar 12g
Final pH7.2 ± 0.2

How to use:
1.Suspend 21g in 1L of distilled or deionized water. Heat with frequent agitation and boil to completely dissolve the powder. Distribute into flasks. Autoclave at 121 for 15 minutes. cooling to 50-55 and adding 40% urea solution.
2.Diluted and treated samples.

Storage: Store in a dark, cool and dry place, tighten the cap immediately after use. Storage period of three years.

500g